UCSC In-Silico PCR
 
>chr12:109099028+109099364 337bp GGGCAGGCACTGTTTTTGTT GCTTAACTCCAGTGTCCCTA
GGGCAGGCACTGTTTTTGTTtgttgttttttaaaagaatttggacataac
atgacaaagaactaatgatgttccaaataacttgcactagaagctttctt
attgaattcttatggtttccaatgagaatctgattttaagtctagtttat
ctttaaatcagctaatgggatttgttgcagaagaaagaaagcattacact
gtttatccacccccacaccaagtcttcacctggacccagatgtgtgacat
aaaagatgtaagtacaacttgttgataatttttattggggagaaggagtc
aaatagtatttttaaatTAGGGACACTGGAGTTAAGC

Primer Melting Temperatures
  Forward: 62.3 C gggcaggcactgtttttgtt
Reverse: 54.5 C gcttaactccagtgtcccta
The temperature calculations are done assuming 50 mM salt and 50 nM annealing oligo concentration. The code to calculate the melting temp comes from Primer3, the formula by Rychlik W, Spencer WJ and Rhoads RE NAR 1990, which can be activated in Primer3 with PRIMER_TM_FORMULA=0.

Help
  What is chr_alt & chr_fix?
Replicating in-Silico PCR results on local machine